Training Purposes Only JEM CERISE JMU (Ver. 1.1) 1 Notice : This technical data is furnished on the condition that it will be used by and disclosed to the receiving Cooperating agency and its contractors and sub contractors only for the purposes of fulfilling the cooperating agency’s responsibilities under the Space Station Intergovernmental Agreement (IGA) and Memorandum of Understanding (MOU). It shall not be used for any other purpose, nor disclosed or retransferred to any other entity or government without prior written permission of the Japan Aerospace Exploration Agency (JAXA). CERISE Experiment “C.elegans RNAi space experiment” JEM CERISE Instructor:Toru Mori Takashi Yamasaki

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 2 Objectives  Perform the CERISE Experiment Operations according to ODFs without instructor’s assistance.  CERISE Experiment Explain the science objectives of the CERISE experiment. List onboard outcomes from performing the CERISE experiment. After this lesson the trainee will be able to:  Operation Flow Outline the CERISE operations flow from the view point of the sample movement.

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 3 Lesson Outline CERISE Experiment Operation Flow of CERISE Key Points for CERISE Operations Appendix 1. Specification of Meas Exp Unit A Schematic of Meas Exp Unit A Photo of Meas Exp Unit A Acronyms

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 4 CERISE Experiment CERISE Experiment (PI: Atsushi Higashitani) Research Objectives 1)Investigate whether RNAi occurs in space as the same manner as on the earth. 2)Explore effects of microgravity on entire gene and protein expressions, and phosphorylations beyond generations. Onboard Outcomes from the Experiment Cultivated nematode Dosimeter Temp Logger Microgravity environment data (taken by MMA) Returned Contained in Culture Bag Holder Downlinked Microgravity Measurement Apparatus (MMA) Culture Bag Holder Returned

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 5 Intentionally Blank

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 6 MELFI Operation Flow of CERISE Saibo Rack/CBEF Activation Meas Exp Unit A (3) Retrieve Saibo Rack/CBEF Deactivation CBEF Cultivation (4 days) -80 deg C Sample Preparation Meas Exp Unit A (4) Instl into CBEF MLT and MMA Deactivation MMA Micro-G environment measurement Sample Retrieve MLT and MMA Activation If they are off As required ULF3(TBD) FLIGHT ULF3 CB Setup CB Stow CB CB Activation Microscope Observation CB Deactivation Microscope Observation :Crew operations :Crew/Ground cooperative operations LEGEND: :Ground operations Meas Exp Unit A (2) Retrieve  Sample Retrieve  Meas Exp Unit A (1) re-installation with dummy mass Cultivation (4 days) Stow Return Meas Exp Unit A Culture Bag Holder ULF3 Launch

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 7 Operation Flow of CERISE (cont’d) Incubator Unit 1G Incubator Unit Micro-G Dummy Mass nematode Culture Bag Holder (4) Pins Remove MELFI (-80 deg C) Sample Preparation Meas Exp Unit A (4) Install Cultivation (4 days) Incubator Unit 1G Incubator Unit Micro-G CBEF nematode Sample Retrieve  Culture Bag x 15  Dosimeter x 1  Temp Logger x 1 Temp Logger Rtrv Stow Culture Bag Microscope Observation Retrieve one Culture Bag From each Attach Observe Stow Meas Exp Unit A Re-Instl with dummy mass Culture Bag Holder (no nematode contained) Install CBEF Re-Instl Cultivation (4 days) Sample Retrieve Microscope Observation Clear Plate CB Microscope Install Reuse Dummy Mass Reuse Stow

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 8 Key Points for CERISE Operations (CBEF) NOTE: Underlined key points have been covered by CBEF Advanced Training. CategoryEquipmentKey Points Crew Safety Electrical ShockSample Tray, Centrifuge Before attaching/detaching the Canisters to/from the Sample Tray/ Centrifuge, turn off power to Canister by switches on the Sample Tray/Centrifuge. Entanglement with rotating device Incubator Unit 1G Verify visually through the Observation Window that Centrifuge is not rotating before opening the door of the Incubator Unit 1G. CentrifugeWhen attaching/detaching the Canisters to/from the 1G Centrifuge, lock turntable of the Centrifuge. Time Critical OperationN/A Dos and Don’tsIncubator UnitDo NOT touch inside Incubator Unit with bare hand to keep it clean. Sample Tray, Centrifuge Be careful not to pinch fingers when releasing drawer locks of slide rails on Sample Tray/Centrifuge. Difficulty in OpsN/A Possible Mis-OpsN/A

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 9 Key Points for CERISE Operations (CB) NOTE: Underlined key points have been covered by CB Advanced Training. CategoryEquipmentKey Points Crew Safety Crew entrapmentAir-tight Chamber When CB is in ‘Stowed Configuration’ fill gaps between the OC and Outer Chassis with Gap Protectors to prevent crew entrapment. Crew Exposure to Broken Glass Operation Chamber Verify no broken glass in Operation Chamber before opening OC door. Time Critical OperationN/A Dos and Don’tsAir-tight Chamber Pull out/stow Air-tight Chamber slowly at about 10 cm per minute to avoid damage to bellows. OC CameraRaise OC Camera (Lens and Flexible Arm) above the inner plate before stowing the Operation Chamber. Difficulty in OpsN/A Possible Mis-OpsN/A

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 10 Appendix 1.Specification of Meas Exp Unit A 2.Schematic of Meas Exp Unit A 3.Photo of Meas Exp Unit A 4.Acronyms

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 11 Specification of Meas Exp Unit A ItemSpecification Size239 x 166 x 102 mm Weight1585 g Power+15V, -15V +12V (for future experiments) Temp Sensor20~45 deg C

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 12 Schematic of Meas Exp Unit A CBEF Circuit Board Analog (A) (0~5V) Analog (B) (0~5V) Temperature Sensor (1) Temperature Sensor (2) +12V +15V -15V NTSC Shield Connector for future experiments Jumper SEL Relay circuit Amplifier circuit

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 13 Overview Inside Photo of Meas Exp Unit A

Training Purposes Only JEM CERISE JMU (Ver. 1.1) 14 Acronyms CB CBEF MELFI MLT MMA OC RNAi Clean Bench Cell Biology Experiment Facility Minus Eighty degree Celsius Laboratory Freezer for ISS MMA Laptop Terminal Microgravity Measurement Apparatus Operation Chamber Ribonucleic Acid Interference